6Y2V

Crystal structure of the double mutant L13R I16K of Low Molecular Weight Protein Tyrosine Phosphatase (LMW-PTP)

6Y2V の概要

• エントリーDOI: 10.2210/pdb6y2v/pdb
• 分子名称: Low molecular weight phosphotyrosine protein phosphatase, TRIETHYLENE GLYCOL, PHOSPHATE ION, ... (4 entities in total)
• 機能のキーワード: engineered, hydrolase
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18306.60

構造登録者

Levy, C. (登録日: 2020-02-17, 公開日: 2020-07-22, 最終更新日: 2024-01-24)

主引用文献

Egbe, E.,Levy, C.W.,Tabernero, L.
Computational and structure-guided design of phosphoinositide substrate specificity into the tyrosine specific LMW-PTP enzyme.
Plos One, 15:e0235133-e0235133, 2020

PubMed Abstract: We have used a combination of computational and structure-based redesign of the low molecular weight protein tyrosine phosphatase, LMW-PTP, to create new activity towards phosphoinositide substrates for which the wild-type enzyme had little or no activity. The redesigned enzymes retain catalytic activity despite residue alterations in the active site, and kinetic experiments confirmed specificity for up to four phosphoinositide substrates. Changes in the shape and overall volume of the active site where critical to facilitate access of the new substrates for catalysis. The kinetics data suggest that both the position and the combination of amino acid mutations are important for specificity towards the phosphoinositide substrates. The introduction of basic residues proved essential to establish new interactions with the multiple phosphate groups in the inositol head, thus promoting catalytically productive complexes. The crystallographic structures of the top-ranking designs confirmed the computational predictions and showed that residue substitutions do not alter the overall folding of the phosphatase or the conformation of the active site P-loop. The engineered LMW-PTP mutants with new activities can be useful reagents in investigating cell signalling pathways and offer the potential for therapeutic applications.

PubMed: 32584877
DOI: 10.1371/journal.pone.0235133

実験手法

X-RAY DIFFRACTION (2 Å)